アプリケーション

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション

Abreviews

特記事項

Immunoelectrophoresis

1/1000.

IHC-FoFr

1/200.

ICC/IF

Use at an assay dependent concentration.

WB

1/1000 - 1/5000. Predicted molecular weight: 10 kDa.

IP

1/100. PubMed: 20103532

追加情報Is unsuitable for IHC-P.

ターゲット情報

関連性Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling.
Similarity: Belongs to the ubiquitin family.
Contains 3 ubiquitin-like domains.

Scrambled and lentivirus-stable infected SN4741 cells grown on coverslips were induced to differentiate under restrictive conditions for 24 h (pre-lethal stage), fixed and permeabilized with 0.1% Triton X-100 + 0.1% sodium citrate for 5 minutes. Cells were blocked with 10% donkey serum in PBS for 1 hour at 37°C and incubated over night at 4°C with rabbit polyclonal antibody against ubiquitin (ab19247, 1:100) and mouse monoclonal α-synuclein antibody (1:25). Samples were washed twice with PBS for 10 minutes before the addition of secondary antibodies at room temperature in the dark for 1 hour

Scrambled infected (control) cells show a diffuse staining pattern and do not show any inclusions at both permissive and restrictive temperatures (upper images). SKP1A-deficient cells developed multiple and perinuclear (bottom images, see arrows and insets) inclusion bodies 24 h after induction of differentiation, with characteristics of aggresomes. The reactivity of the aggregates to the different antibodies demonstrated a similar pattern as indicated by co-localized yellow immunostaining in the overlaid right-hand panel. No fluorescence was detected when the primary antibody was omitted. Each panel shows a representative picture of 10–15 views in two independent experiments.

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Further to correspondence with the source of this antibody I can tell you that the following approach was employed for the testing of this antibody by western blot analysis. In brief the samples were prepared in denaturi...

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It has not been determined whether ab14372 detects mono- or poly-ubiquitinated proteins. It would quite likely recognize both.
Ab19247 should recognize ubiquinated proteins in Western blots regardless of how may ubiq...